Tumor 3'-deoxy-3'-(18)F-fluorothymidine ((18)F-FLT) uptake by PET correlates with thymidine kinase 1 expression: static and kinetic analysis of (18)F-FLT PET studies in lung tumors.

UNLABELLED: We report the first, to our knowledge, findings describing the relationships between both static and dynamic analysis parameters of 3'-deoxy-3'-(18)F-fluorothymidine ((18)F-FLT) PET and the expression of the proliferation marker Ki-67, and the protein expression and enzymatic activity of thymidine kinase-1 (TK1) in surgically resected lung lesions. METHODS: Static and dynamic analyses (4 rate constants and 2 compartments) of (18)F-FLT PET images were performed in a cohort of 25 prospectively accrued, clinically suspected lung cancer patients before surgical resection (1 lesion was found to be benign after surgery). The maximal and overall averaged expression of Ki-67 and TK1 were determined by semiquantitative analysis of immunohistochemical staining. TK1 enzymatic activity was determined by in vitro assay of extracts prepared from flash-frozen samples of the same tumors. RESULTS: Static (18)F-FLT uptake (partial-volume-corrected maximum-pixel standardized uptake value from 60- to 90-min summed dynamic data) was significantly correlated with the overall (ρ = 0.57, P = 0.006) and maximal (ρ = 0.69, P < 0.001) immunohistochemical expressions of Ki-67 and TK1 (overall expression: ρ = 0.65, P = 0.001; maximal expression: ρ = 0.68, P < 0.001) but not with TK1 enzymatic activity (ρ = 0.34, P = 0.146). TK1 activity was significantly correlated with TK1 protein expression only when immunohistochemistry was scored for maximal expression (ρ = 0.52, P = 0.029). Dynamic analysis of (18)F-FLT PET revealed correlations between the flux constant (K(FLT)) and both overall (ρ = 0.53, P = 0.014) and maximal (ρ = 0.50, P = 0.020) TK1 protein expression. K(FLT) was also associated with both overall (ρ = 0.59, P = 0.005) and maximal (ρ = 0.63, P = 0.002) Ki-67 expression. We observed no significant correlations between TK1 enzyme activity and K(FLT). In addition, no significant relationships were found between TK1 expression, TK1 activity, or Ki-67 expression and any of the compartmental rate constants. CONCLUSION: The absence of observable correlations of the imaging parameters with TK1 activity suggests that (18)F-FLT uptake and retention within cells may be complicated by a variety of still undetermined factors in addition to TK1 enzymatic activity.

Hypermethylation of CCND2 May Reflect a Smoking-Induced Precancerous Change in the Lung.

It remains unknown whether tobacco smoke induces DNA hypermethylation as an early event in carcinogenesis or as a late event, specific to overt cancer tissue. Using MethyLight assays, we analyzed 316 lung tissue samples from 151 cancer-free subjects (121 ever-smokers and 30 never-smokers) for hypermethylation of 19 genes previously observed to be hypermethylated in nonsmall cell lung cancers. Only APC (39%), CCND2 (21%), CDH1 (7%), and RARB (4%) were hypermethylated in >2% of these cancer-free subjects. CCND2 was hypermethylated more frequently in ever-smokers (26%) than in never-smokers (3%). CCND2 hypermethylation was also associated with increased age and upper lobe sample location. APC was frequently hypermethylated in both ever-smokers (41%) and never-smokers (30%). BVES, CDH13, CDKN2A (p16), CDKN2B, DAPK1, IGFBP3, IGSF4, KCNH5, KCNH8, MGMT, OPCML, PCSK6, RASSF1, RUNX, and TMS1 were rarely hypermethylated (<2%) in all subjects. Hypermethylation of CCND2 may reflect a smoking-induced precancerous change in the lung.

Synthesis and in vitro evaluation of 5-fluoro-6-[(2-iminopyrrolidin-1-YL)methyl]uracil, TPI(F): an inhibitor of human thymidine phosphorylase (TP).

An investigation was conducted to determine if the 5-fluoro analog of TPI (5-chloro-6-[(2-iminopyrrolidin-1-yl)methyl]uracil), a potent inhibitor of human thymidine phosphorylase (TP), has an IC(50) in a range that might allow to use it labeled for imaging of TP expression in vivo. The previously unreported fluoro analog, TPI(F), was prepared and tested against TPI and TPI(Br) using an inhibition assay of [H-3]thymidine cleavage. An assay, performed in the presence of 0.4 mg/ml of human TP, yielded IC(50) values of 2.5 nM, 2.7 nM, and 9.0 nM for TPI, TPI(Br), and TPI(F), respectively. The results indicate that further studies to develop (18)F-labeled TPI(F) as a potential radiopharmaceutical for PET imaging of TP expression in vivo are warranted.

Reliability and validity of the SmartDiet Canadian version questionnaire.

Measuring the outcomes of nutrition interventions in health promotion settings can be a challenge. Nutrition screening tools are often used to measure dietary patterns and nutrition intervention outcomes, but these tools may lack quantitative scoring methods. Using the SmartDiet questionnaire from Oslo, Norway, as a model, the SmartDiet Canadian Version questionnaire was developed to assess fiber and fat intake. The purpose of this study was to assess the reliability and validity of this new questionnaire. In 2007, 54 volunteer subjects (63% men, mean age 68.3+/-9.7 years) were recruited from the Cardiac Rehabilitation Program at Vancouver General Hospital, Vancouver, BC, Canada. Test-retest reliability was assessed by having subjects complete the questionnaire at recruitment and 1 month later. Intraclass correlation coefficients of reliability (ICC) for both fiber and fat intake were calculated. For fiber scores the ICC was 0.66 (95% confidence interval 0.48, 0.79) and for fat scores the intraclass correlation coefficient was 0.74 (95% confidence interval 0.59, 0.84). Validity was assessed by correlating the completed questionnaire at recruitment with a 3-day food diary. The daily fat and fiber scores from the questionnaire were correlated with the corresponding average daily fat and fiber scores from the 3-day food diary by means of the Spearman correlation coefficient. For fiber scores, Spearman rho was 0.53 (P<0.001), and for fat scores it was 0.48 (P<0.001). The SmartDiet Canadian Version questionnaire is a reliable and valid tool for assessing the daily fiber and fat intake of cardiac rehabilitation clients.

DNA hypermethylation of tumors from non-small cell lung cancer (NSCLC) patients is associated with gender and histologic type.

BACKGROUND: We previously identified a number of genes which were methylated significantly more frequently in the tumor compared to the non-cancerous lung tissues from non-small cell lung cancer (NSCLC) patients. Detection of methylation profiles of genes in NSCLC could provide insight into differential pathways to malignancy and lead to strategies for better treatment of individuals with NSCLC. METHODS: We determined the DNA methylation status of 27 genes using quantitative MethyLight assays in lung tumor samples from 117 clinically well-characterized NSCLC patients. RESULTS: Hypermethylation was detected in one of more of the genes in 106 (91%) of 117 cases and was detected at high levels (percentage methylation reference (PMR)> or =4%) in 79% of NSCLC cases. Methylation of APC, CCND2, KCNH5 and, RUNX was significantly more frequent in adenocarcinomas compared to squamous cell carcinomas (SCC), while methylation of CDKN2A was more common in SCC. Hypermethylation of KCNH5, KCNH8, and RARB was more frequent in females compared to males. Hypermethylation of APC and CCND2 was inversely associated with proliferation score assessed by Ki-67 level. CONCLUSIONS: Our findings of differential gene hypermethylation frequencies in tumor tissues from patients with adenocarcinoma or squamous cell cancers and in females compared to males suggests that further investigation is warranted in order to more fully understand the potential disparate pathways and/or risk factors for NSCLC associated with histologic type and gender.

Thymidine kinase 1 and thymidine phosphorylase expression in non-small-cell lung carcinoma in relation to angiogenesis and proliferation.

The thymidine salvage pathway enzymes thymidine kinase 1 (TK1) and thymidine phosphorylase (TP) compete for thymidine as a substrate and catalyze opposing synthetic and catabolic reactions that have been implicated in the control of proliferation and angiogenesis, respectively. We investigated the relationship between the expression of TK1 and TP as they relate to proliferation (Ki-67 labeling index) and angiogenesis (Chalkley count of CD31-stained blood vessels) in a series of 110 non-small-cell lung cancer (NSCLC) tumors from patients prospectively enrolled in an imaging trial. TK1 and TP exhibited similar patterns of immunohistochemical distribution, in that each was found in both the nucleus and the cytoplasm of tumor cells. Each enzyme exhibited a significant positive correlation between its levels of nuclear and cytoplasmic expression. A significant positive correlation between TK1 expression and the Ki-67 labeling index (r = 0.53, p<0.001) was observed. TP was significantly positively correlated with Chalkley scoring of CD31 staining in high vs low Chalkley scoring samples (mean TP staining of 115.8 vs 79.9 scoring units, p<0.001), respectively. We did not observe a substantial inverse correlation between the TP and TK1 expression levels in the nuclear compartment (r = -0.17, p=0.08). Tumor size was not found to be associated with TK1, TP, Ki-67, or Chalkley score. These findings provide additional evidence for the role of thymidine metabolism in the complex interaction of proliferation and angiogenesis in NSCLC.

Relationship between non-small cell lung cancer FDG uptake at PET, tumor histology, and Ki-67 proliferation index.

INTRODUCTION: We compared primary non-small cell lung cancer (NSCLC) F-fluorodeoxyglucose (FDG) uptake at positron emission tomography (PET) to tumor histologic features and Ki-67 proliferation index. This large, prospectively-recruited patient cohort has previously been analyzed based on differences in FDG uptake across stage groups; the current analysis adds further dimensions to this characterization. MATERIALS AND METHODS: One hundred seventy-eight patients with potentially-resectable NSCLC were scanned with FDG PET before therapy. A partial volume correction algorithm was used to correct FDG uptake values for their dependence on tumor size. Primary tumor resection specimens, core biopsies, and biopsies of metastatic lymph nodes were used to assess each tumor's NSCLC histologic subtype, degree of differentiation, and Ki-67 proliferation index. RESULTS: Bronchioalveolar carcinomas were found to have lower FDG uptake at PET and lower Ki-67 scores than any other histologic subtype. Non-bronchioalveolar adenocarcinomas had lower FDG uptake and Ki-67 scores than squamous cell carcinomas or large cell undifferentiated carcinomas. Better differentiated NSCLCs had lower FDG uptake and Ki-67 scores than more poorly differentiated NSCLCs. There was a significant positive correlation between FDG uptake and Ki-67 scores. Partial volume correction increased the strength of this correlation, while also diminishing the strong positive correlation between FDG uptake and tumor size. CONCLUSIONS: There are significant differences in NSCLC FDG uptake across histologic subtypes and differentiation groups. These differences parallel nearly identical differences in Ki-67 scores, implying that differences in NSCLC tumor cell proliferation may give rise to commensurate differences in tumor glucose metabolism.

DNA methylation in tumor and matched normal tissues from non-small cell lung cancer patients.

We used MethyLight assays to analyze DNA methylation status of 27 genes on 49 paired cancerous and noncancerous tissue samples from non-small cell lung cancer (NSCLC) patients who underwent surgical resection. Seven genes (RARB, BVES, CDKN2A, KCNH5, RASSF1, CDH13, and RUNX) were found to be methylated significantly more frequently in tumor tissues than in noncancerous tissues. Only methylation of CCND2 and APC was frequently detected in both cancerous and noncancerous tissues, supporting the hypothesis that the methylation of these two genes is a preneoplastic change and may be associated with tobacco smoking exposure. Methylation of any one of eight genes (RASSF1, DAPK1, BVES, CDH13, MGMT, KCNH5, RARB, or CDH1) was present in 80% of NSCLC tissues but only in 14% of noncancerous tissues. Detection of methylation of these genes in blood might have utility in monitoring and detecting tumor recurrence in early-stage NSCLC after curative surgical resection.

A simple quantitative assay for the activity of thymidine kinase 1 in solid tumors.

INTRODUCTION: The activity of the pyrimidine salvage pathway enzyme thymidine kinase 1 (TK1) is tightly cell cycle regulated and has been investigated as a prognostic indicator of cancer in a variety of tissues. However, using the in vitro assay of TK1 to rank order a series of unique tumor samples by their TK1 activity can be problematic due to the complex nature of TK1 enzyme substrate kinetics. We present a refined TK1 in vitro assay and method of analysis which address these problems. METHODS: Extracts were prepared of the resected lung lesions from eight patients and assayed for TK1 activity using an in vitro assay modified to account for nonlinearities in extract protein concentration. A separate extract of exponentially growing A549 human lung carcinoma cells was used as a cross-assay control. RESULTS: In extracts prepared from eight frozen samples of resected human lung lesions, TK1 activity (mean=0.0070+/-0.0077 pmol [(3)H]-TMP/microg protein/minute) was 2 orders of magnitude below that of exponentially growing A549 human lung carcinoma cells (mean=0.1572+/-0.0218 pmol [(3)H]-TMP/microg protein/minute; n=9). TK1 activity was nonlinear with respect to extract protein concentration in both groups, with A549 cell extracts exhibiting evidence of positive cooperativity which could not be explained by the presence of detergents in the cell lysis buffer. Lung tumor extracts demonstrated evidence of negative cooperativity. CONCLUSIONS: The modified TK1 assay takes into account these nonlinearities by averaging the results of several complete time-course curves measured over a range of extract protein concentrations. An extract prepared from exponentially growing A549 cells is included in each assay for use as a cross-assay control. We demonstrate that these modifications allow for the accurate rank ordering of TK1 activity in solid tumors.

Toxicology evaluation of radiotracer doses of 3'-deoxy-3'-[18F]fluorothymidine (18F-FLT) for human PET imaging: Laboratory analysis of serial blood samples and comparison to previously investigated therapeutic FLT doses.

BACKGROUND: 18F-FLT is a novel PET radiotracer which has demonstrated a strong potential utility for imaging cellular proliferation in human tumors in vivo. To facilitate future regulatory approval of 18F-FLT for clinical use, we wished to demonstrate the safety of radiotracer doses of 18F-FLT administered to human subjects, by: 1) performing an evaluation of the toxicity of 18F-FLT administered in radiotracer amounts for PET imaging, 2) comparing a radiotracer dose of FLT to clinical trial doses of FLT. METHODS: Twenty patients gave consent to a 18F-FLT injection, subsequent PET imaging, and blood draws. For each patient, blood samples were collected at multiple times before and after 18F-FLT PET. These samples were assayed for a comprehensive metabolic panel, total bilirubin, complete blood and platelet counts. 18F-FLT doses of 2.59 MBq/Kg with a maximal dose of 185 MBq (5 mCi) were used. Blood time-activity curves were generated for each patient from dynamic PET data, providing a measure of the area under the FLT concentration curve for 12 hours (AUC12). RESULTS: No side effects were reported. Only albumin, red blood cell count, hematocrit and hemoglobin showed a statistically significant decrease over time. These changes are attributed to IV hydration during PET imaging and to subsequent blood loss at surgery. The AUC12 values estimated from imaging data are not significantly different from those found from serial measures of FLT blood concentrations (p = 0.66). The blood samples-derived AUC12 values range from 0.232 ng x h/mL to 1.339 ng x h/mL with a mean of 0.802 +/- 0.303 ng x h/mL. This corresponds to 0.46% to 2.68% of the lowest and least toxic clinical trial AUC12 of 50 ng x h/mL reported by Flexner et al (1994). This single injection also corresponds to a nearly 3,000-fold lower cumulative dose than in Flexner's twice daily trial. CONCLUSION: This study shows no evidence of toxicity or complications attributable to 18F-FLT injected intravenously.

Fluorodeoxyglucose uptake of primary non-small cell lung cancer at positron emission tomography: new contrary data on prognostic role.

PURPOSE: This prospective study evaluated the prognostic significance of (18)F-fluorodeoxyglucose ((18)F-FDG) uptake in primary non-small cell lung cancer (NSCLC) at positron emission tomography, in a carefully staged population, while correcting for partial volume effects. EXPERIMENTAL DESIGN: Two hundred eight potentially resectable NSCLC patients were referred for FDG positron emission tomography staging after thoracic computed tomography. Each tumor stage was confirmed surgically, or for some stage IV tumors by additional imaging. The tumor maximum pixel-standardized uptake value (maxSUV) and the maxSUV partial volume corrected for lesion size (PVCmaxSUV) were compared with overall survival and disease-free survival using Cox proportional hazards regression. RESULTS: Stage distribution: stage I, 36%; stage II, 15%; stage III, 30%; stage IV, 19%. Patients were followed for a median of 33.6 months, with 90 deaths from NSCLC (median survival for all stages, 43.3 months). With respect to overall survival, the most significant cutoff value for both maxSUV and PVCmaxSUV was 7. MaxSUV > or =7 was significantly associated with an increased risk of death from NSCLC in univariable analysis, whereas PVCmaxSUV > or =7 was only marginally associated. However, in multivariable analyses, neither maxSUV > or =7 nor PVCmaxSUV > or =7 provided significant additional prognostic information over stage, tumor size, and age. In the 103 patients who underwent surgical resection only, surgical stage, but not maxSUV or PVCmaxSUV, was univariably associated with survival or recurrence. SUV definitions based on lean body mass, body surface area, and plasma glucose correction yielded identical results. CONCLUSIONS: As expected, tumor stage is prognostic in NSCLC. However, tumor FDG uptake does not provide additional prognostic information. This prospective study contradicts prior reports.

Evaluation of 5'-deoxy-5'-[F-18]fluorothymidine as a tracer of intracellular thymidine phosphorylase activity.

Two human cell lines (A549 and U937) with cytosolic thymidine phosphorylase (TP) activity were used to evaluate the potential of 5'-deoxy-5'-[F-18]fluorothymidine ([F-18]DFT) as a tracer of intracellular TP expression. Cellular metabolism of DFT led to the production of 5-[F-18]fluoro-2,5-dideoxy-D-ribose-1alpha-phosphate ([F-18]FddR-1P), in analogy to the metabolism of thymidine, which produces 2-deoxy-D-ribose-1alpha-phosphate (dR-1P). A549 cells showed the highest production rate of FddR-1P. After A549 cells were exposed to [F-18]DFT for 40 min, the relative intracellular concentration of [F-18]FddR-1P was more than sevenfold higher in cells than its precursor in the incubating medium. For the same amount of time, a twofold concentration was seen in U937 cells. However, uptake ratios did not rank with the corresponding TP activities found in cell extracts [TP activity ratio (U937:A549)=1.6] that were independently determined with a labeled thymidine/thymine cleavage assay. The discrepancy of TP activity ratios was traced to the instability of FddR-1P in cells. This was evident from the fact that cells accumulated radioactivity by producing FddR-1P, but activity also effluxed from cells over 1 h when the medium was subsequently made tracer free. The dominant labeled molecule released by cells was characterized as a neutral and lipophilic molecule, which was presumed to be a deoxynucleoside. Our results indicate that [F-18]DFT would not be effective for imaging TP expression because its initial metabolite undergoes further conversion to a diffusible secondary metabolite, allowing activity loss from cells.

Relationship between non-small cell lung cancer fluorodeoxyglucose uptake at positron emission tomography and surgical stage with relevance to patient prognosis.

PURPOSE: Because the tumor stage is the most significant prognostic factor for non-small cell lung cancer (NSCLC) and given that NSCLC [(18)F]fluorodeoxyglucose ((18)F-FDG) uptake appears to have prognostic significance, we examined the relationship between NSCLC (18)F-FDG uptake and surgical stage. EXPERIMENTAL DESIGN: One hundred seventy-eight patients with a proven diagnosis of NSCLC were enrolled, then imaged with (18)F-FDG positron emission tomography and their disease thoroughly staged. Primary tumor size at computed tomography and (18)F-FDG uptake were compared to overall tumor stage and to T, N, and M stage descriptors. Tumor uptake was quantitated by maximum pixel-standardized uptake value (maxSUV) and then partial volume corrected for lesion size using recovery coefficients. RESULTS: A significant difference in tumor size was associated with tumors of different TNM stage, T status, N status, or M status. Similarly, the primary tumor maxSUV was significantly associated with TNM stage, T status, and M status. However, we observed no significant difference in the partial-volume-corrected tumor maxSUV for different stages; different T, N, or M descriptors; tumors without evidence of spread (N(0)M(0)) versus tumors with nodal spread (N(1,2,3)M(0)); or tumors without spread (N(0)M(0)) versus all others. CONCLUSIONS: We found an association between tumor stage and (18)F-FDG maxSUV, but this relationship disappeared after correction of tumor uptake for lesion size. Therefore, if partial-volume-corrected (18)F-FDG uptake is prognostic of NSCLC outcome, it is not on the basis of a relationship with tumor stage but through a different mechanism.

Application of a neural network to improve nodal staging accuracy with 18F-FDG PET in non-small cell lung cancer.

UNLABELLED: We proposed to train a back-propagation artificial neural network (aNN) on a cohort of surgically proven non-small cell lung cancers (NSCLCs) and compare its accuracy with that of a trained (18)F-FDG PET reader. We plan to show that an aNN trained on (18)F-FDG PET- and CT-derived data is more accurate in predicting the true surgicopathologic nodal stage than a human reader. METHODS: One hundred thirty-three NSCLC patients with surgically proven N status treated at the University of Washington Medical Center or the Veterans Affairs Puget Sound Health Care System between February 1998 and September 2002 were used as inputs for the creation of an aNN. From CT of the thorax and (18)F-FDG PET (neck to pelvis) performed before surgery, we extracted the primary tumor size and uptake (maximum pixel SUV [maxSUV]), normal lung and mediastinal uptake, and nodal uptake (maxSUV). Using the same 133 cases, the same output (surgical N status, N(0) to N(3)), and the same software configuration settings, scenarios were created to assess which input parameters were most influential in creating an optimal aNN. To compute this optimal aNN, cases were split randomly 100 times into a training subset of 103 cases and a testing subset of 30 cases having the same proportion of N(0), N(1), N(2), and N(3) cases. N status predicted by the aNN was compared with the proven surgical N status to calculate the aNN accuracy. The N status readings from (18)F-FDG PET were also compared with the surgical N status for the same cases to determine (18)F-FDG PET accuracy. RESULTS: Statistical tests demonstrate that the best aNN accuracy is achieved by using N(1)-N(2)- N(3) nodal maxSUV divided by background uptake, the primary tumor size, and primary tumor maxSUV as inputs. The aNN correctly predicted the N stage in 87.3% of the testing cases compared with 73.5% for the (18)F-FDG PET expert reader. Accuracy of the aNN increased to 94.8% (PET, 89.4%) when comparing N(0) + N(1) with N(2) or N(3) status and to 94.9% (PET, 91.9%) when comparing N(0) + N(1) with N(2) + N(3) status. CONCLUSION: A back-propagation aNN can be trained to predict hilar and mediastinal nodal involvement with greater accuracy than an expert (18)F-FDG PET reader. Such a tool could be used to improve clinical interpretations and for clinical training.

Validation of FLT uptake as a measure of thymidine kinase-1 activity in A549 carcinoma cells.

UNLABELLED: The thymidine analog (18)F-3'-deoxy-3' -fluorothymidine (FLT) is being used clinically for PET imaging of tumor proliferation. Appropriate use of this tracer requires validating the mechanisms by which it accumulates in dividing cells. We tested the accuracy with which FLT uptake predicted the activity of cytosolic thymidine kinase-1 (TK(1)), an enzyme that is upregulated before and during DNA synthesis. METHODS: Cultured A549 human lung carcinoma cells were manipulated to a range of proliferation rates from actively dividing to growth arrested. Uptake of radiolabeled FLT was compared with cell cycle activity, which was expressed as the percentage of cells in S phase, and with activity of cytosolic TK(1). We also compared uptake of FLT and deoxyglucose. We genetically manipulated A549 cells by transfecting them with human papillomavirus type 16 E6 (designated A549-E6) to abrogate function of the tumor suppressor gene, p53. Although radiation typically inhibits progression of mammalian cells through the cell cycle, abrogation of p53 function eliminates this inhibition. We then compared FLT uptake with the percentage of cells in S phase and TK(1) activity in irradiated A549-E6 cells and in irradiated control cells having normal p53 function and the expected radiation-induced growth delay. RESULTS: A549 cells with only 3%-5% cells in S phase took up little FLT and had low levels of TK(1) activity. When cells were stimulated to grow by being placed into fresh medium, we observed a strong correlation between increased FLT uptake and increased TK(1) activity. As expected, FLT uptake varied much more as a function of growth than did uptake of deoxyglucose. Nonproliferating A549 cells did not enter the cell cycle if they were irradiated before being placed into fresh medium, and they did not accumulate FLT or show elevated TK(1) activity. In contrast, radiation did not inhibit the cell cycle progression of A549-E6 cells. When subcultured, they began to grow and showed increased uptake of FLT commensurate with greater TK(1) activity. CONCLUSION: In cultured A549 cells FLT uptake is positively correlated with cell growth and TK(1) activity. Inhibition of cell cycle progression prevents FLT uptake and increased TK(1) activity. These results suggest that FLT images reflect TK(1) activity and the percentage of cells in S phase.